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Method for accurate detection of whole genome methylation sites by utilizing trace genome DNA (deoxyribonucleic acid)

A whole-genome and genome-wide technology, applied in the field of solexa sequencing technology and second-generation sequencing technology, can solve the problems of trace DNA sample research, low PCR amplification efficiency, etc., and achieve the effect of increasing PCR amplification efficiency and reducing damage

Active Publication Date: 2012-04-11
BGI TECH SOLUTIONS
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Problems solved by technology

It overcomes the shortcomings of inability to mix samples, low PCR amplification efficiency and inability to study trace DNA samples in conventional methylation sequencing

Method used

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  • Method for accurate detection of whole genome methylation sites by utilizing trace genome DNA (deoxyribonucleic acid)
  • Method for accurate detection of whole genome methylation sites by utilizing trace genome DNA (deoxyribonucleic acid)
  • Method for accurate detection of whole genome methylation sites by utilizing trace genome DNA (deoxyribonucleic acid)

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Embodiment Construction

[0050] Embodiments of the present invention will be described in detail below in conjunction with examples, but those skilled in the art will understand that the following examples are only used to illustrate the present invention, and should not be considered as limiting the scope of the present invention.

[0051] Using the method of the present invention, we used 30ng and 100ng of human peripheral blood whole genome DNA (genomic DNA extracted from the blood of a Chinese adult male) as starting materials to construct two trace whole genome methylation high-throughput sequencing libraries. The quality of the two libraries was tested by sanger sequencing method, and 100ng of the libraries were subjected to high-throughput whole-genome sequencing (Illumina GA). Simultaneously compared with 100ng people's whole genome DNA of peripheral blood (the genomic DNA extracted from the blood of a Chinese adult male) as the starting material using the method of micro-library construction o...

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Abstract

According to the invention, based on Illumina common tag library sequencing and methylation normal sequencing and combining a common library tag sequencing method, a new method for accurate detection of whole genome methylation sites by utilizing trace genome DNA (deoxyribonucleic acid) is built. According to the method, in the construction of a methylation library by using trace genome DNA, exogenous DNA is innovatively added to carry out bisulfite high-efficiency co-treatment; and at the same time, fragment size selection is not needed, and PCR (polymerase chain reaction) amplification is directly carried out after bisulfite treatment. By the method, the defects that in the common methylation sequencing, samples can not be mixed, the PCR amplification efficiency is low and the trace DNAsample can not be researched are overcome.

Description

technical field [0001] The invention relates to the technical field of high-throughput sequencing of methylation, in particular to the technical field of high-throughput sequencing of trace DNA whole genome methylation. In addition, the present invention also relates to a tag sequencing technology and a method for constructing a tag library in the same reaction system for multiple samples. The method of the present invention is particularly suitable for the second generation sequencing technology, especially the solexa sequencing technology. Background technique [0002] DNA methylation is the most deeply studied epigenetic mechanism. DNA methylation plays an important role in maintaining normal cell function, inhibiting the damage of genome integrity caused by parasitic DNA components, modifying chromatin structure, inactivating X chromosome, genome imprinting, embryo It plays an important role in development and human tumorigenesis, and is one of the new research hotspots...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C40B50/06C40B40/08C12Q1/68C12N15/11
CPCC40B40/08C12Q1/6827C40B50/06C12N15/1093
Inventor 孙继华闫淑静王君文罗慧娟
Owner BGI TECH SOLUTIONS
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